Fluorescent dyes for multiphoton bio-imaging applications

Katherine J. Schafer; Sheng Yao; Kevin D. Belfield; Joel M. Hales; David J. Hagan; Eric W. Van Stryland

doi:10.1117/12.529412

14 June 2004 Fluorescent dyes for multiphoton bio-imaging applications

Katherine J. Schafer, Sheng Yao, Kevin D. Belfield, Joel M. Hales, David J. Hagan, Eric W. Van Stryland

Proceedings Volume 5329, Genetically Engineered and Optical Probes for Biomedical Applications II; (2004) https://doi.org/10.1117/12.529412
Event: Biomedical Optics 2004, 2004, San Jose, CA, United States

Abstract

Fluorescent dyes and probes are key components in multiphoton based fluorescence microscopy imaging of biological samples. While many commercially available fluorescent dyes have sufficed, most exhibit relatively low two-photon absorption (2PA) cross-section values in the tunability range of Ti:sapphire lasers commonly used in multiphoton microscopy imaging. Furthermore, available fluorophores may be plagued with either low fluorescence quantum yields and/or the additional problem of rapid photobleaching upon exposure to the high peak powers provided by fs laser sources. In order to address the demand for better performing dyes for two-photon based imaging, we have prepared a new series of reactive fluorophores tailored for multiphoton imaging. These fluorophores are based upon the fluorene ring system, known to exhibit high fluorescence quantum yields, typically > 0.7, and possess high photostability. They have been functionalized with moieties to act, e.g., as efficient amine-reactive fluorescent probes for the covalent attachment onto, e.g., proteins and antibodies. The synthesis and the single-photon spectral characteristics, as well as measured two-photon absorption cross sections of the reactive fluorophores in solution will be presented. Spectral characterizations of bovine serum albumin (BSA) conjugated with the new reactive probe will also be presented.

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Katherine J. Schafer, Sheng Yao, Kevin D. Belfield, Joel M. Hales, David J. Hagan, and Eric W. Van Stryland "Fluorescent dyes for multiphoton bio-imaging applications", Proc. SPIE 5329, Genetically Engineered and Optical Probes for Biomedical Applications II, (14 June 2004); https://doi.org/10.1117/12.529412

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KEYWORDS

Luminescence

Absorption

Proteins

Quantum efficiency

Absorbance

Multiphoton microscopy

Spectroscopy

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